Analysis of PCR-Based VNTR Markers to Evaluate Engraftment Status after Bone Marrow Transplantation / 대한임상병리학회지

ABSTRACT

BACKGROUND: The evaluation of engraftment after BMT may be effectively accomplished by the analysis of genomic polymorphism, such as variable number of tandem repeat (VNTR). Discrimination potential (PD) and allelic profile of VNTR locus might be varied widely between races and geographic areas. Thus PCR-based VNTR loci to establish test panel useful in evaluating engraftment status of Korean patients after BMT were analyzed. MATERIAL AND METHODS: Thirty normal adults (15 males and 15 females), and each patient with acute lymphoblastic leukemia and severe aplastic anemia who had undergone allogeneic BMT were tested. Genomic DNAs extracted from peripheral blood lymphocytes or hair follicles were subjected to three PCR long tandem repeats (LTRs) and fifteen PCR short tandem repeats (STRs) loci analysis using silver-stain mode of detection. RESULTS: The PCR sensivity of VNTR system tested, and detection limit of minor component in mixing experiment, were 100 pg and 0.1%, respectively. The most informative marker was ACTBP2 with 93.2% of PD, and 98.0% of actual PD (APD). The most informative test panel was ACTBP2, D3S2386 and D1S1768 loci-combination with 99.6% of PD and 100.0% of combined APD. CONCLUSIONS: STRs, especially combination of ACTBP2, D3S2386, and D3S11768, were thought to be very useful screening markers for evaluating engraftment status in nonsibling allogeneic BMT. But most of allogeneic BMT are carried out between siblings, who have similar genetic marker each other, so further evaluation is need in sibling-BMT.